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Identification of the natural ligand of an orphan G-protein-coupled receptor involved in the regulation of vasoconstriction

Nature Cell Biology volume 1, pages 383–385 (1999)Cite this article

Homology-based cloning approaches and genome-sequencing efforts have revealed the existence of a large number of human genes encoding ‘orphan’ G-protein-coupled receptors (GPCRs), receptors that bind unidentified natural ligands. Discovery of these natural ligands is the first necessary step in understanding the biological significance of the orphan GPCRs. We1 and others2 have developed an approach by which to successfully isolate endogenous ligands from complex tissue libraries. This approach, referred to as the orphan-receptor strategy3, uses orphan receptors as baits to isolate their native ligand from tissue extracts and has been successfully applied to identify new neuropeptides4,5,6. Here we apply this strategy to the orphan receptor GPR14 and show that it binds the bioactive peptide known as urotensin II.

As GPR14 is similar to the somatostatin receptors we decided to screen somatostatin-like, cysteine-bridge-containing peptides on GPR14 and to compare their biological activities and physicochemical properties with that of the endogenous compound identified in our purification scheme. Peptides tested were human melanin-concentrating hormone (MCH), somatostatin-14 (SST-14) and cortistatin-14 and urotensin II. Whereas MCH was inactive, SST-14 and the related peptide cortistatin activated GPR14, albeit only at very high concentrations (Table 1). In contrast, urotensin II induced a robust increase in [Ca2+]i, with a median effective concentration (EC50) of 0.14 nM (Table 1), showing a potency more than 25,000-fold greater than that of SST-14.

Table 1 Pharmacological profiles of GPR14 and SSTR2A.
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Figure 1: Identification of urotensin II as the endogenous ligand of GPR14.

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Acknowledgements

This work was supported by a grant from the NIH and by the endowment of the Eric and Lila Nelson Chair in Neuropharmacology. We thank D. Knauer for help with iodinations, R. Reinscheid for editorial assistance and S. Lin for helpful discussions.

Correspondence and requests for materials should be addressed to O.C.

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Authors and Affiliations

  1. Department of Pharmacology, University of California at Irvine, Irvine, 92697-4625, California, USA

    Hans-Peter Nothacker, Zhiwei Wang, Anne Marie McNeill, Yumiko Saito, Sven Merten, Sue P. Duckles & Olivier Civelli

  2. Department of Pharmacology and Medicine, University of Toronto, Toronto, M5S 2S1, Ontario, Canada

    Brian O’Dowd

  3. Department of Developmental and Cell Biology, University of California at Irvine, Irvine, 92697-4625, California, USA

    Olivier Civelli

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  1. Hans-Peter Nothacker
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Correspondence to Olivier Civelli.

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Nothacker, HP., Wang, Z., McNeill, A. et al. Identification of the natural ligand of an orphan G-protein-coupled receptor involved in the regulation of vasoconstriction. Nat Cell Biol 1, 383–385 (1999). https://doi.org/10.1038/14081

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